title: Liquid chromatography-tandem mass spectrometry detection of the quaternary ammonium compound mebezonium as an active ingredient in t61.
abstract: Quaternary ammonium compounds pose an analytical challenge. Mebezonium, a muscle-relaxing agent contained in veterinary euthanasia solution T61, was analyzed in body fluids, organs, and injection sites of a veterinarian by liquid chromatography-tandem mass spectrometry (LC-MS-MS) method. Additionally, embutramide and tetracaine, which are two other active ingredients contained in T61, methadone, xylazine, and analgesics were detected by LC-MS-MS and high-performance liquid chromatography-ultraviolet detection methods. For detection of mebezonium a solid-phase extraction (SPE) combined with ionpairing reagent heptafluorobutyric acid was developed. Separation was achieved on Phenomenex Synergi Hydro RP C(18) column combined with ammonium formate buffer and acetonitrile (pH 3.5). To enrich other drugs, liquid-liquid extraction procedures were used. Most of these drugs were separated on a Restek Allure PFP Propyl column using the mentioned mobile phase. Mebezonium and embutramide were detected in femoral vein serum in concentrations of 10.9 and 2.0 mg/L, respectively. The concentration of xylazine and methadone in serum was 2.0 and 0.4 mg/L, respectively. The LC-MS-MS method with SPE combined with an ion-pairing reagent allowed the quantitation of mebezonium. Methadone was detected in toxic concentrations and was, in combination with xylazine and T61, considered to be the cause of death.
authors.affiliation: Institute of Forensic Medicine, University of Bonn, Bonn, Germany.
authors.forename: Katrin M
authors.initials: KM
authors.lastname: Kirschbaum
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Grellner
authors.forename: Gertrud
authors.initials: G
authors.lastname: Rochholz
authors.forename: Frank
authors.initials: F
authors.lastname: Musshoff
authors.forename: Burkhard
authors.initials: B
authors.lastname: Madea
mesh_terms: D058647:Adrenergic alpha-2 Receptor Agonists / Q000737:chemistry / Q000378:metabolism; D000577:Amides / Q000737:chemistry* / Q000378:metabolism; D000700:Analgesics / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D004338:Drug Combinations; D053593:Forensic Toxicology; D006801:Humans; D006993:Hypnotics and Sedatives / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D000644:Quaternary Ammonium Compounds / Q000737:chemistry* / Q000378:metabolism; D013405:Suicide; D053719:Tandem Mass Spectrometry; D013748:Tetracaine / Q000737:chemistry* / Q000378:metabolism; D019155:Veterinary Drugs / Q000737:chemistry / Q000378:metabolism; D014991:Xylazine / Q000737:chemistry / Q000378:metabolism
chemical_list: D058647:Adrenergic alpha-2 Receptor Agonists; D000577:Amides; D000700:Analgesics; D004338:Drug Combinations; D006993:Hypnotics and Sedatives; D000644:Quaternary Ammonium Compounds; D019155:Veterinary Drugs; D013748:Tetracaine; D014991:Xylazine; C015591:T 61; C059325:mebezonium